Yeast Protocols

ISBN-13: 9781493907984 / Angielski / Twarda / 2014 / 313 str.

Wei Xiao

Opis

Yeast Protocols, Third Edition presents up-to-date advances in research using yeasts as models. Chapters cover topics such as basic protocols in yeast culture and genomic manipulation, protocols that study certain organelles such as mitochondria and peroxisomes and their functions in autophagy and assays commonly used in yeast-based studies that can be adapted to other organisms. As the first sequenced living organism, budding yeast S. cerevisiae and other model yeasts have helped greatly in life science research. The easy switch between the haploid and diploid state makes yeast a paradigm of genetic manipulation. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and notes on troubleshooting and avoiding known pitfalls.Authoritative and easily accessible, Yeast Protocols, Third Edition seeks to serve both professionals and novices with newly-developed protocols to study this essential model organism.

Spis treści

1. Basic Investigations in Saccharomyces cerevisiae

            Brendan Curran and Virginia Bugeja

2. Isolation of Yeast Nucleic Acids

            Michael Biss, Michelle Hanna and Wei Xiao

3. Quantification of GFP Signals by Fluorescent Microscopy and Flow Cytometry

            Ting Wei and Heping Dai

4. Yeast Transformation by the LiAc/SS Carrier DNA/PEG Method

            R. Daniel Gietz

5. Targeted Gene Deletion in Saccharomyces cerevisiae and Schizosaccharomyces pombe

            Johannes H. Hegemann, Sven Boris Heick, Jennifer Pöhlmann, Markus Langen and Ursula Fleig

6. See & Catch Method for Studying Protein Complexes in Yeast Cells: A Technique Unifying Fluorescence Microscopy and Mass Spectrometry

            Changhui Deng and Andrew N. Krutchinsky

7. Yeast Two Hybrid Liquid Screening

            Elisa Donnard, Erica M. Queiroz, J. Miguel Ortega, and R. Daniel Gietz

8. Targeted Mutagenesis of a Specific Gene in Yeast

            Likui Zhang, Alina Radziwon, and Linda J. Reha-Krantz

9. Fluorescence Staining of Mitochondria for Morphology Analysis in S. cerevisiae

            Kai Stefan Dimmer

10. Assays for Autophagy I: the Cvt Pathway and Non-Selective Autophagy

            Wei-Pang Huang, Takahiro Shintani, and Zhiping Xie

11. Assays for Autophagy II: Mitochondrial Autophagy

            Tomotake Kanki and Koji Okamoto

12. Biochemically Characterizing the Subcellular Localization of Peroxisomal Proteins by Fractionation, Protease Protection and Carbonate Extraction

            Danielle Hagstrom and Changle Ma

13. Yeast Survival and Growth Assays

            Xin Xu, Amanda Lambrecht, and Wei Xiao

14. Spontaneous Mutagenesis Assay

            Susan Blackwell, Michelle Hanna, and Wei Xiao

15. Mutational Specificity Analysis: Assay for Mutations in the Yeast SUP4-o Gene

            Bernard A. Kunz

16. piggyBac Transposon-Based Insertional Mutagenesis for the Fission Yeast Schizosaccharomyces pombe

            Jun Li and Li-Lin Du

17. Replicative and Chronological Life Span Assays

            Spike D. L. Postnikoff and Troy A. A. Harkness

18. Detection of Protein Arginine Methylation in Saccharomyces cerevisiae

            Christopher Aa. Jackson and Michael C. Yu

19. Detection of Protein Post-translational Modifications from Whole Cell Extracts in Saccharomyces cerevisiae

            Lindsay G. Ball and Wei Xiao

20. Ligand Engineering Using Yeast Surface Display

            Benjamin J. Hackel

21. Screening Combinatorial Libraries of Cyclic Peptides Using the Yeast Two-Hybrid Assay

            Kris Barreto and C. Ronald Geyer